Neoplastic cells carry upon their surfaces antigens which distinguish them from their normal counterparts, with chemically induced neoplasms frequently expressing unique antigens, denoted tumor specific transplantation antigens (TSTA). The applicant has demonstrated that TSTA activity can be extracted from intact tumor cells using single-phase solutions of n-butanol. The extraction technique is not cytotoxic, as judged by the ability of extracted cells to proliferate in vitro and in vivo. Crude butanol extracts are immunogenic in immunoprotection assays and evoke a positive delayed hypersensitivity(DH) response in immune mice. However, crude butanol extracts do not contain measurable amounts of alloantigenic activity. In addition to butanol, the applicant has shown that low concentrations of Octylglucoside also removes immunogenic TSTA activity from intact cells without affecting viability. Thus, two noncytolytic extraction techniques have been developed for the preparation of TSTA moieties which are stable in aqueous media. This application proposes isolation and purification of the TSTA molecules from a panel of 3-MCA-induced fibrosarcomas of C3H/HeJ mice. The butanol extracted materials will be fractionated by isoelectric focusing, molecular sieve chromatography, polyacrylamide gel electrophoresis, hydroxyapatite chromatography and affinity chromatography. The presence of an active TSTA moiety will be assessed by the induction of immunoprotection and by evocation of a DH response. During purification, the applicant will characterize the TSTA active principles with respect to charge, apparent size, association with lipids and degree of glycosylation. The third objective of the proposal is to study the kinetics of antigen synthesis and turnover following extraction and the influence of the cell cycle on antigen expression. The de novo expression of TSTA upon the surface of butanol extracted cells will be monitored by immunoprecipitation of pulse-labeled antigen. The rate of decline in the amount of labeled antigen will reflect the rate of antigen turnover. Comparison of the rates of antigen synthesis and turnover in untreated and extracted cells may elucidate extraction-related alterations in membrane protein metabolism. Finally, the ability of extracted MCA-F cells to absorb an anti-MCA-F antiserum will be a measure of the re-expression of surface molecules following extraction.